The Salt-Sensitive Structure and Zinc Inhibition of Borrelia Burgdorferi Protease BbHtrA

Mol Microbiol. 2016 Feb;99(3):586-96. doi: 10.1111/mmi.13251. Epub 2015 Nov 19.

Abstract

HtrA serine proteases are highly conserved and essential ATP-independent proteases with chaperone activity. Bacteria express a variable number of HtrA homologues that contribute to the virulence and pathogenicity of bacterial pathogens. Lyme disease spirochetes possess a single HtrA protease homologue, Borrelia burgdorferi HtrA (BbHtrA). Previous studies established that, like the human orthologue HtrA1, BbHtrA is proteolytically active against numerous extracellular proteins in vitro. In this study, we utilized size exclusion chromatography and blue native polyacrylamide gel electrophoresis (BN-PAGE) to demonstrate BbHtrA oligomeric structures that were substrate independent and salt sensitive. Examination of the influence of transition metals on the activity of BbHtrA revealed that this protease is inhibited by Zn(2+) > Cu(2+) > Mn(2+). Extending this analysis to two other HtrA proteases, E. coli DegP and HtrA1, revealed that all three HtrA proteases were reversibly inhibited by ZnCl2 at all micro molar concentrations examined. Commercial inhibitors for HtrA proteases are not available and physiologic HtrA inhibitors are unknown. Our observation of conserved zinc inhibition of HtrA proteases will facilitate structural and functional studies of additional members of this important class of proteases.

Publication types

  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Bacterial Proteins / chemistry*
  • Bacterial Proteins / genetics
  • Bacterial Proteins / metabolism*
  • Borrelia burgdorferi / chemistry
  • Borrelia burgdorferi / enzymology*
  • Borrelia burgdorferi / genetics
  • Borrelia burgdorferi / metabolism
  • Chlorides / chemistry
  • Chlorides / metabolism*
  • Enzyme Inhibitors / chemistry
  • Enzyme Inhibitors / metabolism*
  • Humans
  • Kinetics
  • Lyme Disease / microbiology
  • Serine Endopeptidases / chemistry*
  • Serine Endopeptidases / genetics
  • Serine Endopeptidases / metabolism*
  • Zinc / chemistry
  • Zinc / metabolism*
  • Zinc Compounds / chemistry
  • Zinc Compounds / metabolism*

Substances

  • Bacterial Proteins
  • Chlorides
  • Enzyme Inhibitors
  • Zinc Compounds
  • zinc chloride
  • Serine Endopeptidases
  • Zinc